Fig 2: SOCS3 expression in A549.A549 cells were treated with DMSO vehicle control or 30 µg/mL H. perforatum extract in DMSO, with or without inoculation of H1N1 influenza virus. SOCS3 mRNA expression levels relative to and GAPDH (control) 3 hrs and 24 hrs after treatment are shown as log relative transcription level (Panel A. Mean ± SEM, N=3). Protein expression of SOCS3 24 hrs after the treatment was measured using Western blot and shown in Panel B (Mean ± SEM, N=3) as percentage of vehicle control without virus, together with a representative image. Values at the same time point are differentiated by individual letter labels when significant differences exist (p<0.05, a > b > c).

Fig 3: Erlotinib inhibits STAT3 phosphorylation of and upregulates SOCS3 expression. HepG2.2.15 cells were cultivated with an equal concentration (0.125%) of DMSO and with or without 10 μM erlotinib for 1 h. STAT3, p-STAT3 and SOCS3 were detected using Western blot analysis. The optical density of interesting bands was showed as the ratio against that of the control of β-actin. (A) HepG2.2.15 cells had much higher levels of STAT3 and p-STAT3 and erlotinib inhibited STAT3 phosphorylation without affection on STAT3 level. (B) Erlotinib increased SOCS3 expression.

Fig 4: HEL cells treated with different concentration of curcumin (0‐30 µmol/L) for 24 h. A, The indicated proteins of JAK2/STAT5 signalling were detected by SDS‐PAGE. Representative Western blots were shown (n = 3). The relative intensity of each band is shown under each blot as fold change (FC) compared to untreated control. B, The mRNA expression levels of PIM family members, Socs‐1 and Socs‐3 were evaluated by qRT‐PCR. Results are expressed as −∆∆Ct (n = 7). Statistical analyses were performed using the two‐tailed Mann‐Whitney Utest, comparing conditions two by two respect to not treated condition. All the analysis with confidence level major of 95% are indicated like significant and marked as followed: *P ≤ 0.05; **P ≤ 0.01;

Fig 5: RNA transcription profiles the lung of BALB/c mice.Expression levels of SOCS3 relative to GAPDH in lungs collected at the end of the 5-day high viral titer inoculation study (107.9 EID50, N=15 for inoculated groups and N=6 for non-inoculated group) and 10-day low viral tier inoculation study (105.1 EID50, N=13 for each group) were measured and shown as log relative transcription level (Mean ± SEM). Values at the same time point are differentiated by individual over bar labels when significant differences were found (p<0.05). ND=not determined (no non-inoculated group in the 10 day study).